Systems and methods for aptamer-based intracellular delivery of a payload using nanoneedles

Abstract

Methods, chips and systems are provided for the ex vivo delivery of payloads into cells using an aptamer-based approach. Methods are provided for delivering a payload to a cell comprising providing a nanoneedle and a polynucleotide, wherein a first end of the polynucleotide comprises an aptamer capable of binding a molecule endogenous to a cell, wherein the first end of the polynucleotide is conjugated to the nanoneedle, and wherein a second end of the polynucleotide comprises an oligonucleotide capable of hybridizing with a payload; contacting the payload with the polynucleotide, wherein the payload contains a nucleotide sequence that is complementary to the oligonucleotide sequence; and inserting the nanoneedle into a cell, wherein upon insertion of the nanoneedle into the cell the payload is released from the polynucleotide.

What is claimed: 
     
         1 . A method of delivering a payload to a cell comprising
 a. providing a nanoneedle and a polynucleotide,
 wherein a first end of the polynucleotide comprises an aptamer capable of binding a molecule endogenous to a cell, 
 wherein the first end of the polynucleotide is conjugated to the nanoneedle, and 
 wherein a second end of the polynucleotide comprises an oligonucleotide capable of hybridizing with a payload; 
   b. contacting the payload with the polynucleotide, wherein the payload contains a nucleotide sequence that is complementary to the oligonucleotide sequence; and   c. inserting the nanoneedle into a cell, wherein upon insertion of the nanoneedle into the cell the payload is released from the polynucleotide.   
     
     
         2 . The method of  claim 1  wherein the aptamer is capable of binding to adenosine triphosphate. 
     
     
         3 . The method of  claim 1  wherein the payload is a nucleotide-based molecule. 
     
     
         4 . The method of  claim 1 , wherein the payload comprises genetic material capable of stably integrating into the genome of the cell. 
     
     
         5 . The method of  claim 1 , wherein the payload comprises genetic material capable of transiently expressing a desired target protein. 
     
     
         6 . The method of  claim 1 , wherein the payload inhibits expression of a target gene. 
     
     
         7 . The method of  claim 1 , wherein the payload is capable of functionally inhibiting a protein. 
     
     
         8 . The method of  claim 1 , wherein the payload comprises genetic material that expresses a protein capable of functionally inhibiting a desired target. 
     
     
         9 . The method of  claim 1 , wherein the payload is a protein. 
     
     
         10 . The method of  claim 1 , wherein the payload is a small molecule. 
     
     
         11 . The method of  claim 1 , wherein the payload is conjugated to a unique nucleotide sequence capable of hybridizing with the oligonucleotide. 
     
     
         12 . The method of  claim 1 , further comprising inserting the nanoneedle into an inner volume of a well that is configured to house the cell, wherein the nanoneedle is configured such that, upon insertion of the nanoneedle into the cell, the payload is released from the polynucleotide. 
     
     
         13 . A chip for delivering a payload to a cell, the chip comprising
 a solid support, and   a nanoneedle attached to the solid support and configured to receive a polynucleotide,
 wherein a first end of the polynucleotide comprises an aptamer capable of binding a molecule endogenous to a cell, 
 wherein the first end of the polynucleotide is capable of conjugating to the nanoneedle, and 
 wherein a second end of the polynucleotide comprises an oligonucleotide capable of hybridizing with one of a plurality of molecules of a payload. 
   
     
     
         14 . The chip of  claim 13 , wherein the chip is comprised of silicon. 
     
     
         15 . The chip of  claim 13 , wherein the aptamer is capable of binding to adenosine triphosphate. 
     
     
         16 . The chip of  claim 13 , wherein the payload is a nucleotide-based molecule. 
     
     
         17 . The chip of  claim 13 , wherein the payload comprises genetic material capable of stably integrating into the cells genome. 
     
     
         18 . The chip of  claim 13 , wherein the payload comprises genetic material capable of transiently expressing a desired target protein. 
     
     
         19 . The chip of  claim 13 , wherein the payload inhibits expression of a target gene. 
     
     
         20 . The chip of  claim 13 , wherein the payload is capable of functionally inhibiting a protein. 
     
     
         21 . The chip of  claim 13 , wherein the payload comprises genetic material that expresses a protein capable of functionally inhibiting a desired target. 
     
     
         22 . The chip of  claim 13 , wherein the payload is a protein. 
     
     
         23 . The method of  claim 13 , wherein the payload is a small molecule. 
     
     
         24 . The method of  claim 13 , wherein the payload is conjugated to a unique nucleotide sequence capable of hybridizing with the oligonucleotide. 
     
     
         25 . The method of  claim 13 , further comprising a plurality of nanoneedles attached to the solid support, wherein each of the plurality of nanoneedles is configured to receive a polynucleotide. 
     
     
         26 . A system for delivering a payload comprising of a plurality of molecules to a cell comprising
 a. a plurality of nanoneedles, and a plurality of polynucleotides,
 wherein a first end of a respective polynucleotide of a respective one of the plurality of polynucleotides comprises an aptamer capable of binding a molecule endogenous to the cell, 
 wherein the first end of the respective polynucleotide is conjugated to one of the plurality of nanoneedles, and 
 wherein a second end of the respective polynucleotide comprises an oligonucleotide capable of hybridizing with one of a plurality of molecules of a payload; 
   b. a plurality of wells; and   c. an injecting device that houses the plurality of nanoneedles thereon, and is configured to move the plurality of nanoneedles to within the plurality of wells.   
     
     
         27 . The system of  claim 25 , wherein the aptamer is capable of binding to adenosine triphosphate. 
     
     
         28 . The system of  claim 25 , wherein the payload is a nucleotide based molecule. 
     
     
         29 . The system of  claim 25 , wherein the payload comprises genetic material capable of stably integrating into the cells genome. 
     
     
         30 . The system of  claim 25 , wherein the payload comprises genetic material capable of transiently expressing a desired target protein. 
     
     
         31 . The system of  claim 25 , wherein the payload inhibits expression of a target gene. 
     
     
         32 . The system of  claim 25 , wherein the payload is capable of functionally inhibiting a protein. 
     
     
         33 . The system of  claim 35 , wherein the payload comprises genetic material that expresses a protein capable of functionally inhibiting a desired target. 
     
     
         34 . The system of  claim 25 , wherein the payload is a protein. 
     
     
         35 . The system of  claim 25 , wherein the payload is a small molecule. 
     
     
         36 . The system of  claim 25 , wherein the payload is conjugated to a unique nucleotide sequence capable of hybridizing with the oligonucleotide. 
     
     
         37 . The system of  claim 25 , wherein the each of the plurality of wells is configured to house a cell to be penetrated by a nanoneedle. 
     
     
         38 . The system of  claim 25 , wherein the injecting device is configured to move the plurality of nanoneedles to within the plurality of wells.